title: Cell-cycle analysis of fission yeast cells by flow cytometry.
abstract: The cell cycle of the fission yeast, Schizosaccharomyces pombe, does not easily lend itself to analysis by flow cytometry, mainly because cells in G(1) and G(2) phase contain the same amount of DNA. This occurs because fission yeast cells under standard growth conditions do not complete cytokinesis until after G(1) phase. We have devised a flow cytometric method exploiting the fact that cells in G(1) phase contain two nuclei, whereas cells in G(2) are mononuclear. Measurements of the width as well as the total area of the DNA-associated fluorescence signal allows the discrimination between cells in G(1) and in G(2) phase and the cell-cycle progression of fission yeast can be followed in detail by flow cytometry. Furthermore, we show how this method can be used to monitor the timing of cell entry into anaphase. Fission yeast cells tend to form multimers, which represents another problem of flow cytometry-based cell-cycle analysis. Here we present a method employing light-scatter measurements to enable the exclusion of cell doublets, thereby further improving the analysis of fission yeast cells by flow cytometry.
authors.affiliation: Department of Cell Biology, Institute for Cancer Research, Oslo University Hospital, Oslo, Norway.
authors.forename: Jon Halvor Jonsrud
authors.initials: JH
authors.lastname: Knutsen
authors.forename: Idun Dale
authors.initials: ID
authors.lastname: Rein
authors.forename: Christiane
authors.initials: C
authors.lastname: Rothe
authors.forename: Trond
authors.initials: T
authors.lastname: Stokke
authors.forename: Beáta
authors.initials: B
authors.lastname: Grallert
authors.forename: Erik
authors.initials: E
authors.lastname: Boye
mesh_terms: D002453:Cell Cycle / Q000235:genetics / Q000502:physiology*; D018797:Cell Cycle Proteins / Q000235:genetics; D049109:Cell Proliferation; D005434:Flow Cytometry / Q000379:methods; D016193:G1 Phase / Q000235:genetics; D008027:Light; D008938:Mitosis / Q000235:genetics / Q000502:physiology; D008954:Models, Biological; D030781:Organisms, Genetically Modified; D012542:Scattering, Radiation; D012568:Schizosaccharomyces / Q000166:cytology* / Q000254:growth & development / Q000502:physiology; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics; D020666:ras-GRF1 / Q000235:genetics
chemical_list: D018797:Cell Cycle Proteins; D029702:Schizosaccharomyces pombe Proteins; D014157:Transcription Factors; C061178:cdc10 protein, S pombe; C080926:cdc22 protein, S pombe; D020666:ras-GRF1